This proposal from UC Davis, Toronto Centre for Phenogenomics (TCP), and Charles River Laboratories, together known as the "DTC" consortium, seeks to establish and operate a Knock-Out Mouse Program Phase 2 (K0MP2) Production Center. The purpose of our Production Center will be to generate 500 deletion mutant mouse lines from IKMC ES cells each year, conduct preliminary analyses on and cryopreserve germplasm from both deletion and conditional-ready mutants, and make mice and data readily available to phenotyping centers and to the research community through IKMC repositories. Our long-term objective is to develop and implement production strategies that increase throughput to facilitate production of up to ~1,250 mutant mouse lines annually at lower cost while ensuring product quality and reliability. Together, our track record of handling and converting ES cells to mice, quality control, testing and analyses, germplasm cryopreservation and recovery, and rapid expansion of breeding colonies, especially on the C57BL/6N background, fully supports our plan to produce all 500 mutant mouse lines per year. Our participation in K0MP1 will enable us to immediately begin to deliver high quality, reliable, and consistent product to the phenotyping centers. Briefly, UCD shall initiate the production pipeline and expand and verify all mutant IKMC ES cells. Fully qc'd ES cells will be converted into chimeras for breeding to germline confirmed mouse lines, excision of the neo cassette and/or critical exon as necessary, lacZ analysis, and cryopreservation of germplasm from conditional alleles. UCD and TCP will then ship heterozygous mice expressing the post- CRE excision deletion allele to CRL for large-scale breeding and expansion, homozygous viability, fertility, and fecundity testing, and embryo cryopreservation. Upon request, CRL will be prepared to ship frozen embryos, breeding pairs, or full cohorts to K0MP2 Phenotyping Centers. UCD, TCP, and CRL shall also deposit mice and germplasm back to the KOMP Repository at UCD for archiving and distribution. In addition, UCD and TCP will conduct several resource-related development projects to enhance handling, production, and analysis of mutant mouse lines. RELEVANCE: Genetically altered mice serve as excellent research models to understand human disease. This study will add significant knowledge to our understanding of the roles of genes in disease processes. With this knowledge, we can better identify causes, treat, and design prevention strategies against diseases, leading to a healthier national population.